Novel promoter

Abstract

The present invention relates a promoter capable of strongly expressing a gene linked downstream (3′ side) of the promoter in prokaryote, especially in Escherichia Coli . The promoter of the invention is a promoter for prokaryote having a novel nucleotide sequence between any −35 region sequence and any −10 region sequence. The promoter has a strong promoter activity and thus a target peptide or protein can be produced at a high efficiency and in large amounts through linking the structural gene encoding the target peptide or protein downstream of the promoter.

1 . A promoter comprising the same or substantially the same nucleotide sequence as that shown by SEQ ID NO: 34.  
     
     
         2 . The promoter according to  claim 1 , which comprises the same or substantially the same nucleotide sequence as that shown by SEQ ID NO: 34 between any type of −35 region and any type of −10 region.  
     
     
         3 . The promoter according to  claim 2 , in which the −35 region has the nucleotide sequence shown by SEQ ID NO: 37.  
     
     
         4 . The promoter according to  claim 2 , in which the −10 region has the nucleotide sequence shown by SEQ ID NO: 38 or 39.  
     
     
         5 . The promoter according to  claim 2 , which has the nucleotide sequence shown by SEQ ID NO: 35.  
     
     
         6 . The promoter according to  claim 2 , which has the nucleotide sequence shown by SEQ ID NO: 36.  
     
     
         7 . A DNA comprising the promoter according to any one of  claims 1  to  6 .  
     
     
         8 . A recombinant vector comprising the promoter according to any one of  claims 1  to  6  or the DNA according to  claim 7 .  
     
     
         9 . The recombinant vector according to  claim 8 , which comprises a DNA having a structural gene to be expressed under the control of the promoter according to any one of  claims 1  to  6 .  
     
     
         10 . The recombinant vector according to  claim 9 , referred to as pNP3GHNO12, in which the structural gene is the human gorwth hormone gene.  
     
     
         11 . A transformant transformed with the recombinant vector according to  claim 9 .  
     
     
         12 . The transformant  Escherichia coli  MM294/pNP3GHNO12 (FERM BP-7611), which is transformed with the the recombinant vector according to  claim 10 .  
     
     
         13 . A method of producing a protein or a salt thereof, which comprises culturing the transformant according to  claim 11  to produce the protein encoded by the structural gene.  
     
     
         14 . A method of producing the human growth hormone or a salt thereof, which comprises culturing the transformant according to  claim 12  to produce the human growth hormone.  
     
     
         15 . The recombinant vector according to  claim 9 , in which the structural gene is a reporter gene.  
     
     
         16 . The recombinant vector according to  claim 15 , in which the reporter gene is kanamycin resistance gene or GFP gene.  
     
     
         17 . A transformant transformed with the recombinant vector according to  claim 15 .  
     
     
         18 . A transformant transformed with the recombinant vector according to  claim 16 .  
     
     
         19 . A DNA comprising the same or substantially the same nucleotide sequence as that shown by SEQ ID NO: 34.  
     
     
         20 . The DNA according to  claim 20 , which comprises the same or substantially the same nucleotide sequence as that shown by SEQ ID NO: 34 between any type of −35 region and any type of −10 region.  
     
     
         21 . Use of the DNA according to  claim 19  or  20  for the production of a promoter.  
     
     
         22 . Use of the promoter according to any one of  claims 1  to  6  for the production of a protein encoded by a structural gene.